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We narrowed to 676 results for: RON

Showing: 241 - 260 of 676 results
  1. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

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    Blog Post
    ...species, such as the human gut bacteria B. thetaiotaomicron, Mycobacterium, Pseudomonas, and E. coli. ...CRISPRi in many types of bacteria: lab-adapted, environmental isolated, and disease-associated. Mobile-CRISPRi...non-model bacteria including pathogenic strains and environmental isolates. For example, Peters and colleagues... remained constant, with fitness effects more pronounced at 12 doublings than at 6. Key takeaways of Mobile-CRISPRi...
  2. Fluorescent Tagging of Endogenous Genes with SapTrap

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    Blog Post
    ...mitochondrial fission/fusion, nuclear import, and neuronal trafficking. Although FPs have enabled discovery...tagging endogenous proteins with an auxin-inducible degron (AID) tag to generate proteins which can be rapidly...Protein Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors.” Cell Rep.15... Michelle Cronin...
  3. Hot Plasmids and Viral Preps - May 2021

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    Blog Post
    ... to combat severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus may acquire mutations...CRISPR/Cas9 constructs. zCas9i is a high efficiency intron-optimized SpCas9-coding gene. SlugCas9 is a compact...selectively in Parvalbumin containing cortical interneurons. We now provide ready-to-use AAV vectors expressing...this promoter. Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f are now available as a...
  4. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

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    Blog Post
    ...many researchers remain unaware that EGFP/GFP is prone to forming noncovalent dimers. This property of ... matter of concentration Most FPs in nature are prone to dimerization (i.e. EGFP) [1, 2] or even form ... fused to the POI. These structures are large (microns), bright, non-physiological, and difficult to ignore...CFP and YFP dimers can form in the absence of environmental changes that the biosensor is designed to detect...
  5. Which Fluorescent Protein Should I Use?

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    Blog Post
    ...change their excitation/emission behavior upon environmental changes (e.g. pH, Ca2+ flux, etc). The most ...Oligomerization: The first generations of FPs were prone to oligomerize. This may affect the biological function... these FPs cannot be used in oxygen deprived environment. Recently, a new GFP isolated from the Unagi ...you are planning to express the FP in acidic environments (e.g. yeast cytosol, which is slightly acidic...
  6. Technical Design of a Western Blot

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    Blog Post
    ...either DTT or β-mercaptoethanol (BME). DTT is a stronger reducing agent than BME and has a less… distinctive...life when stored properly. Because BME is not as strong as DTT, you’ll have to use different concentrations...quantifying your blot. CCD cameras have a much higher upfront cost, but require no consumables, produce digital...like Coomassie Blue or Ponceau S. It is far less prone to error than normalizing to so-called housekeeping...happy blotting! Many thanks to Addgenies Ashley Waldron, Meghan Rego, and Amrita Rhodes for their help ...
  7. CRISPR 101: Any Base Transversion Editors

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    Blog Post
    ...refresher if you ever lose track of which acronym is which. Acronym Full name Notes   Base transition ...close at hand? Once again, this post has a lot of acronyms, so we've included a glossary at the end of this... for saturation mutagenesis, and they provide a strong proof of concept for further development. ...
  8. Hot Plasmids - November 2023

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    Blog Post
    ...experimental systems. They demonstrated LOV-Turbo in neurons, in pulse-chase experiments, and more. Find LOV-Turbo...element (TRE) or the cFos promoter. For example, neurons treated with a phorbol ester activator gave fluorescent...Images of iPAK4 fibers in cultured rat hippocampal neurons. HaloTag dyes are switched from yellow to red at... Teaching an old antibody new tricks By Ashley Waldron While IHC protocols typically call for thin sections...
  9. Viral Production at Addgene

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    Blog Post
    ...Boyden (Addgene viral prep # 84445-AAVrg).   Electron Microscopy The ratio of empty to full (i.e., genome-containing...representative vector preparations was quantified with electron microscopy after negative staining. Empty vector...than full vector particles.     Figure 3: Electron micrograph of AAV vector preparation shows that...psPAX2 and pMD2.G packaging system developed by the Trono lab. Cell culture medium (OptiPro SFM) containing...
  10. AAV Titers: Where Do They Come From And What Do They Mean?

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    Blog Post
    ...DNA dot blot...Transduction assay...SDS-PAGE...Electron microscopy… Any guesses? That's right... ways...genome-containing viral capsids is typically quantified by electron microscopy of a viral vector solution. Since this... this technique is intensive and requires an electron microscope, it is not routinely performed on all...genome-containing (Figure 1).    Figure 1: Electron micrograph of an Addgene AAV viral vector preparation...
  11. CRISPR Activation: A Practical Guide

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    Blog Post
    ...pluripotent stem cells (Chavez et al., 2015) into neuronal cells by activating genes that cause differentiation...CRISPR-based activation can be made inducible upon environmental triggers. This is done by creating a split dCas9...CRISPR activation are also reversible: once the environmental trigger leaves, gene expression is no longer...Dow D, Buser-Doepner C, Bertotti A, Trusolino L, Stronach EA, Saez-Rodriguez J, Yusa K, Garnett MJ (2019...
  12. Plasmids 101: Broad Host Range Plasmids

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    Blog Post
    ...contains iterons that serve as recognition sites for replication initiation proteins. These iterons vary ...plasmid with multiple oris that vary in DnaA-box and iteron structure, host-independent   Building broad...Development of constitutive and IPTG-inducible integron promoter-based expression systems for Escherichia...
  13. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR/Cas

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    Blog Post
    ... methods.   Rapid detection of 2019 novel coronavirus SARS-CoV-2 using a CRISPR based DETECTR assay...in the saliva (Lucia et al., 2020). Detecting coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​...ultrasensitive and visual detection of novel coronavirus SARS-CoV-2 Sensitivity: 4.6-11 copies/microliter...2020). An ultrasensitive, rapid, and portable coronavirus SARS-CoV-2 sequence detection method based on...
  14. Multiple Plasmids at a Low Price: Kits from Addgene

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    Blog Post
    ...reports 5 (2015). BIOFAB plasmid set 3000 Series Confronted with the problem of creating vectors for predictable...Shine-Delgarno sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can ...of varying strengths centered within GFP/RFP bicistronic reporters characterized in Cambray et al., 2013...terminators can be used to precisely tune your polycistronic E. coli expression constructs. Some applications...
  15. Troubleshooting and Optimizing a Western Blot

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    Blog Post
    ...aggregation in your prep. If your proteins are the wrong size on your gel or blot, Table 1 (at the end of...and keep an eye on it and the leading line (dye front) visible in the sample buffer. You’ll want to run...interest, but you don’t want to run so long your dye front or ladder runs off the gel. Luckily, this is easily...easily monitored in real time, as the dye front (and many ladders!) are visible to the naked eye.   If ... Troubleshooting Suggestion Proteins are the wrong size Proteins may be aggregating Lyse at lower...
  16. Deep Dive: Statistical Tests (Comparisons)

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    Blog Post
    ...to reduce false positives within a test.   The Bonferroni Correction and the Benjamini-Hochberg procedure...control for large numbers of comparisons. The Bonferroni does this by establishing a significance threshold...However, if many comparisons need to be performed, Bonferroni and Benjamini will greatly reduce family-wise...Dependent Variables: A Monte Carlo Simulation Study. Front. Appl. Math. Stat. Volume 2 (2016). doi.org/10.3389...
  17. Illuminating Choices: A Guide to Selecting Fluorescent Dyes and Ligands

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    Blog Post
    ...Heinrichs, 2009). When a fluorophore absorbs light, electrons are elevated from a ground state to an excited...After a brief elevation in this excited state, the electron returns to its ground state by emitting a photon...Pyrgiotakis, G., Zhang, Z., Purington, E., Gramlich, W., Grondin, Y., Rogers, R., Bousfield, D., & Demokritou, P...nanocellulose for nanotoxicological studies. Environmental Science: Nano, 6(5), 1516–1526. https://doi....
  18. With an Eye Towards the Future, We Look Back at the March for Science

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    Blog Post
    ...many scientists operate in a hyper-competitive environment where getting hired hinges on performing near...government scientists to the de-funding of crucial environmental research programs, Canada has been here before...researcher in the University of Michigan Department of Astronomy. She studies how planets form around young stars...community of science supporters is much larger and stronger that I expected. Jennifer Stancill, Ph.D., is ...
  19. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

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    Blog Post
    ... Studying cell mobility within these complex environments has never been easy and, for many years, researchers...same laser) cooperate to cause a higher-energy electronic transition in a fluorescent molecule (see figure...adaptive and innate immunity in diverse tissue environments. These studies provide quantitative measurements...-parasite interactions in the brains of mice chronically infected with Toxoplasma gondii." The Journal...
Showing: 241 - 260 of 676 results